Screening method for identifying anti-PACAP antibodies or antibody fragments suitable for use in treating or preventing PACAP-associated photophobia or light aversion

Abstract

This invention relates to methods of screening for anti-PACAP antibodies, or anti-PACAP receptor antibodies, and antigen binding fragments thereof, for potential use in treating or preventing PACAP-associated photophobia or light aversion, and therapeutic compositions containing and methods of using anti-PACAP antibodies, or anti-PACAP receptor antibodies, and antigen binding fragments thereof.

What is claimed is: 
     
       1. A method of screening for an antibody or antigen binding fragment suitable for use in treating or preventing Pituitary Adenylate Cyclase-Activating Peptide (PACAP)-associated photophobia or light aversion, or precluding the onset of PACAP-associated photophobia or light aversion, in a subject in need thereof,
 which method comprises: 
 (i) providing at least one first test subject and at least one second test subject; 
 (ii) administering PACAP to the at least one first test subject and the at least one second test subject; 
 (iii) further administering to the at least one first test subject one or more anti-PACAP antibodies, or an antigen binding fragment of any of the foregoing, wherein said one or more antibodies or antigen binding fragments includes one comprising a variable heavy chain polypeptide having the heavy chain CDRs1-3 of SEQ ID NO: 44, 46 and 48 respectively; and a variable light chain polypeptide having the light chain CDRs1-3 of SEQ ID NO: 64, 66 and 68 respectively; or one comprising the variable heavy chain polypeptide of SEQ ID NO: 42 and the variable light chain polypeptide of SEQ ID NO: 62; 
 (iv) comparing the response of the at least one first test subject and the at least one second test subject to light; and 
 (v) based on this comparison, identifying one or more antibodies or antigen binding fragments thereof that yield a decreased light aversion or decreased photophobia in the at least one first test subject as compared with the at least one second test subject, thereby identifying an antibody or antigen binding fragment suitable for use in treating or preventing PACAP-associated photophobia or light aversion, or precluding the onset of PACAP-associated photophobia or light aversion in a subject in need thereof; or 
 which method comprises: 
 (i) providing at least one first test subject and at least one second test subject; 
 (ii) administering to the at least one first test subject one or more anti-PACAP antibodies, or an antigen binding fragment of any of the foregoing, wherein said one or more antibodies or antigen binding fragments thereof includes one comprising a variable heavy chain polypeptide having the heavy chain CDRs1-3 of SEQ ID NO: 44, 46 and 48 respectively; and a variable light chain polypeptide having the light chain CDRs1-3 of SEQ ID NO: 64, 66 and 68 respectively; or one comprising the variable heavy chain polypeptide of SEQ ID NO: 42 and the variable light chain polypeptide of SEQ ID NO: 62; 
 (iii) administering PACAP to the at least one first test subject and the at least one second test subject; 
 (iv) comparing the response of the at least one first test subject and the at least one second test subject to light; and 
 (v) based on this comparison, identifying antibodies or antigen binding fragments that yield decreased photophobia or decreased light aversion in the at least one first test subject as compared with the at least one second test subject, identifying an antibody or antigen binding fragment suitable for use in treating or preventing PACAP-associated photophobia or light aversion, or identifying an antibody or antigen binding fragment suitable for precluding the onset of PACAP-associated photophobia or light aversion, in a subject to be treated in need thereof. 
 
     
     
       2. The method of  claim 1 , further comprising one or more of the following:
 (i) further confirming in a human subject, the efficacy of the one or more antibodies or antigen binding fragments for inhibiting PACAP-associated photophobia or light aversion, or precluding the onset of PACAP-associated photophobia or light aversion; 
 (ii) the at least one first test subject and/or the at least one second test subject is a mammal; 
 (iii) the at least one first test subject and/or at least one second test subject is (i) a mouse, monkey, rabbit, human, rat, guinea pig, dog, or hamster, wherein optionally the monkey is a macaque, marmoset, tamarin, spider monkey, owl monkey, vervet monkey, squirrel monkey, or baboon; or (ii) a mouse, optionally a CD1 mouse; 
 (iv) the identified antibody or antigen binding fragment thereof specifically binds PACAP; and 
 (v) the method further comprises adapting the identified antibody or antigen binding fragment for use in treating a subject who suffers from one or more of migraine, hemiplegic migraines, cluster headaches, migrainous neuralgia, chronic headaches, tension headaches, secondary headaches due to an underlying structural problem in the head or neck, cranial neuralgia, sinus headaches, allergy-induced headaches, headache, or other migraine condition. 
 
     
     
       3. The method of  claim 1 , wherein
 (i) the subject to be treated has an ocular disorder associated with photophobia selected from one or more of achromatopsia, aniridia, photophobia caused by an anticholinergic drug, aphakia (absence of the lens of the eye), buphthalmos (abnormally narrow angle between the cornea and iris), cataracts, cone dystrophy, congenital abnormalities of the eye, viral conjunctivitis (“pink eye”), corneal abrasion, corneal dystrophy, corneal ulcer, disruption of the corneal epithelium, ectopia lentis, endophthalmitis, eye trauma caused by disease, injury, infection, optionally chalazion, episcleritis, glaucoma, keratoconus, optic nerve hypoplasia, hydrophthalmos, congenital glaucoma iritis, optic neuritis, pigment dispersion syndrome, pupillary dilation (naturally or chemically induced), retinal detachment, scarring of the cornea or sclera, and uveitis; or 
 (ii) the subject to be treated has a nervous-system-related or neurological condition associated with photophobia selected from one or more of autism spectrum disorders, chiari malformation, dyslexia, encephalitis including myalgic encephalomyelitis (chronic fatigue syndrome), meningitis, subarachnoid hemorrhage, tumor of the posterior cranial fossa, ankylosing spondylitis, albinism, ariboflavinosis, long term use of benzodiazepines, withdrawal from benzodiazepines, chemotherapy, chikungunya infection, cystinosis, Ehlers-Danlos syndrome, hangover, influenza infection, infectious mononucleosis, magnesium deficiency, mercury poisoning, migraine, rabies, and tyrosinemia type II (Richner-Hanhart syndrome); or 
 (iii) the subject to be treated has a photophobia-associated disorder selected from one or more of migraine (with or without aura), iritis, uveitis, meningitis, depression, bipolar disorder, cluster headache or another trigeminal autonomic cephalalgia or blepharospasm, depression, post-traumatic stress syndrome (PTSD) traumatic brain injury, and agoraphobia; or 
 (iv) the at least one first subject and/or the at least one second subject suffers from migraine headaches. 
 
     
     
       4. The method of  claim 1 , wherein
 (i) the one or more antibodies or antigen binding fragments thereof are for use in combination with another active agent for treating migraine, or are for use as a monotherapy; 
 (ii) the anti-PACAP antibody or fragment thereof comprises:
 an immunoglobulin variable heavy chain having the CDR1 sequence of SEQ ID NO: 44; a CDR2 sequence of SEQ ID NO: 46; and a CDR3 sequence of SEQ ID NO: 48; and an immunoglobulin variable light chain having the CDR1 sequence of SEQ ID NO: 64; a CDR2 sequence of SEQ ID NO: 66; and a CDR3 sequence of SEQ ID NO: 68; and/or 
 
 (iii) the antibody or fragment thereof is a humanized antibody or fragment thereof; 
 (iv) the antibody or fragment thereof is a chimeric antibody or fragment thereof; 
 (v) the antibody or fragment thereof comprises a single chain antibody or fragment thereof; 
 (vi) the chimeric antibody or fragment thereof comprises a human Fc, optionally a human Fc is derived from IgG1, IgG2, IgG3, or IgG4; 
 (vii) the anti-PACAP antibody or fragment thereof binds to PACAP with a binding affinity (K D ) of less than or equal to 5×10 −5  M, 10 −5  M, 5×10 −6  M, 10 −6  M, 5×10 −7  M, 10 −7  M, 5×10 −8  M, 10 −8  M, 5×10 −9  M, 10 −9  M, 5×10 −10  M, 10 −10  M, 5×10 −11  M, 10 −11  M, 5×10 −12  M, 10 −12  M, 5×10 −13  M, or 10 −13  M, as determined by ELISA, bio-layer interferometry (“BLI”), kinetics exclusion assay, or surface plasmon resonance at 25° C. or 37° C.; 
 (viii) the anti-PACAP antibody or fragment thereof binds to PACAP with a KD that is less than about 100 nM, less than about 40 nM, less than about 1 nM, less than about 100 pM, less than about 50 pM, or less than about 25 pM; 
 (ix) the anti-PACAP antibody or fragment thereof binds to PACAP with a KD that is between about 10 pM and about 100 pM; 
 (x) the antibody or fragment thereof is entirely non-glycosylated, or lacks N-glycosylation, or contains only mannose residues; 
 (xi) the antibody or fragment thereof contains an Fc region that is modified to alter effector function, half-life, proteolysis, and/or glycosylation; 
 (xii) the anti-PACAP antibody or fragment thereof specifically binds to circulating soluble PACAP molecules in vivo; 
 (xiii) the anti-PACAP antibody or fragment thereof specifically binds to PACAP27 and/or PACAP38; 
 (xiv) the affinity of said anti-PACAP antibody or fragment thereof for PACAP is at least 10-fold, 30-fold, 100-fold, 300-fold, 1000-fold, 3000-fold, 10000-fold, 30000-fold, 100000-fold, 300000-fold, 1000000-fold, 3000000-fold, 10000000-fold, 30000000-fold or more stronger than the affinity of said anti-PACAP antibody and antigen binding fragment to VIP; 
 (xv) the anti-PACAP antibody or fragment thereof (a) inhibits or neutralizes at least one biological effect elicited by PACAP; (b) neutralizes or inhibits PACAP activation of at least one of PAC1-R, VPAC1-R and/or VPAC2-R; (c) neutralizes or inhibits PACAP activation of each of PAC1-R, VPAC1-R and VPAC2-R; (d) neutralizes or inhibits PACAP activation of PAC1-R; (e) is capable of inhibiting PACAP binding to at least one of PAC1-R, VPAC1-R and/or VPAC2-R; (f) is capable of inhibiting PACAP binding to each of PAC1-R, VPAC1-R and/or VPAC2-R; (g) is capable of inhibiting PACAP binding to PAC1-R; and/or (h) inhibits PACAP-induced cAMP production; 
 (xvi) the anti-PACAP antibody or fragment thereof inhibits the association of PACAP with one or more PACAP receptors including PAC-1R, VPAC1-R, and/or VPAC2-R; 
 (xvii) the antibody or fragment thereof is administered intramuscularly, subcutaneously, intravenously, rectally, by infusion, orally, transdermally, or by inhalation; 
 (xviii) the antibody or fragment thereof is administered intravenously; 
 (xix) the antibody or fragment thereof is administered with an additional therapeutic agent or regimen selected from anti-histamines, anti-inflammatory agents, and antibiotics; 
 (xx) the antibody or fragment thereof is directly or indirectly attached to a detectable label or therapeutic agent; or 
 (xxi) the antibody or fragment thereof further comprises an effector moiety optionally a detectable moiety or a functional moiety, further optionally wherein the detectable label is a fluorescent dye, an enzyme, a substrate, a bioluminescent material, a radioactive material, or a chemiluminescent material and the functional moiety is streptavidin, avidin, biotin, a cytotoxin, a cytotoxic agent, or a radioactive material. 
 
     
     
       5. A method of assessing the potential in vivo efficacy of one or more candidate anti-Pituitary Adenylate Cyclase-Activating Peptide (PACAP) antibodies, or antigen binding fragments thereof, for treating PACAP-associated photophobia or light aversion, comprising determining whether the one or more antibodies or fragments thereof inhibit or diminish light aversion behavior in a first rodent administered PACAP, as compared to a second rodent administered PACAP in the presence of said one or more candidate anti-PACAP antibodies or fragments thereof, wherein said one or more anti-PACAP antibodies or fragments thereof comprise an anti-PACAP antibody or antigen binding fragment thereof which comprises a variable heavy chain polypeptide having the heavy chain CDRs1-3 of SEQ ID NO: 44, 46 and 48 respectively; and a variable light chain polypeptide having the light chain CDRs1-3 of SEQ ID NO: 64, 66 and 68 respectively; or comprise the variable heavy chain polypeptide of SEQ ID NO: 42 and the variable light chain polypeptide of SEQ ID NO: 62. 
     
     
       6. The method of  claim 5 , wherein this assay is used to assess whether the one or more anti-PACAP antibodies or fragments thereof may be effective for treatment of a neurological condition characterized by increased PACAP levels. 
     
     
       7. The method of  claim 5 , wherein this assay is used to assess whether the one or more anti-PACAP antibodies or fragments thereof may be effective for treatment of migraine, menstrual migraine, or chronic migraine. 
     
     
       8. The method of  claim 5 , wherein this assay is used to assess whether the one or more anti-PACAP antibodies or fragments thereof may be effective for treatment of migraines (with or without aura. 
     
     
       9. The method of  claim 5 , which further comprises administering an active agent to said first and/or second rodent selected from: ibuprofen, naproxen, sumatriptan, paracetamol/acetaminophen, caffeine, a triptan, a corticosteroid, and combinations thereof.